Simultaneous detection of three fish rhabdoviruses using multiplex real-time quantitative RT-PCR assay
| Autor: | Gao Longying, Yong Teng, Feiyan Tian, Yulin Jiang, Junqiang He, Congxin Xie, Zongxiao Liu, Xiayang Xie, Huifang Li, Hong Liu, Jingshun Yang, Jiangqiang Lv, Shi Xiujie |
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| Rok vydání: | 2007 |
| Předmět: |
Infectious hematopoietic necrosis virus
viruses Sensitivity and Specificity Virus law.invention Cell Line Novirhabdovirus Fish Diseases law Virology Rhabdoviridae Infections Multiplex polymerase chain reaction Animals Multiplex Carp Infectious pancreatic necrosis virus Polymerase chain reaction biology Reverse Transcriptase Polymerase Chain Reaction Fishes Vesiculovirus biology.organism_classification Molecular biology Viral hemorrhagic septicemia |
| Zdroj: | Journal of virological methods. 149(1) |
| ISSN: | 0166-0934 |
| Popis: | Spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV) are three important fish rhabdoviruses, causing serious Office International des Epizooties (OIE) classified diseases in wild and farmed fish. Here, a new multiplex real-time quantitative RT-PCR (mqRT-PCR) assay was developed for simultaneous detection, identification and quantification of these three rhabdoviruses. The sets of primers and probes were targeted to conserved regions of glycoprotein (G) gene of SVCV, nucleoprotein (N) gene of IHNV and G gene of VHSV and used to amplify. The sensitivity, specificity and interference test of mqRT-PCR assay was analyzed. It was shown that the detection levels of 100 copies of SVCV, 220 copies of IHNV and 140 copies of VHSV were achieved, and there was no non-specific amplification and cross-reactivity using RNA of pike fry rhabdovirus (PFRV), infectious pancreatic necrosis virus (IPNV) and grass carp reovirus (GCRV). A total of 80 clinical fish samples were tested using the mqRT-PCR assay and the results were confirmed by antigen-capture ELISA and cell culture assay. This assay has the potential to be used for both research applications and diagnosis. |
| Databáze: | OpenAIRE |
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